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Literature summary extracted from

  • Feng, L.; Sun, Y.; Deng, H.; Li, D.; Wan, J.; Wang, X.; Wang, W.; Liao, X.; Ren, Y.; Hu, X.
    Structural and biochemical characterization of fructose-1,6/sedoheptulose-1,7-bisphosphatase from the cyanobacterium Synechocystis strain 6803 (2014), FEBS J., 281, 916-926.
    View publication on PubMed

Cloned(Commentary)

EC Number Cloned (Comment) Organism
3.1.3.37 expressed in Escherichia coli BL21(DE3) cells Synechocystis sp.

Crystallization (Commentary)

EC Number Crystallization (Comment) Organism
3.1.3.11 bifunctional D-fructose 1,6-bisphosphatase/sedoheptulose 1,7-bisphosphatase in complex with AMP and fructose 1,6-bisphosphate, to 2.3 A resolution. The allosteric inhibitor AMP and the substrate fructose 1,6-bisphosphate exhibit an unusual binding mode when in complex with D-fructose 1,6-bisphosphatase/sedoheptulose 1,7-bisphosphatase. AMP binds to the allosteric sites near the interface across the up/down subunit pairs C1C4 and C2C3 in the center of the tetramer, while fructose 1,6-bisphophate binds opposite to the interface between the horizontal subunit pairs C1C2 or C3C4. Formation of a disulfide linkage may occur between Cys75 and Cys99. The enzyme may be regulated through ligand binding and alteration of the structure of the enzyme complex Synechocystis sp.
3.1.3.37 hanging drop vapor diffusion method, using 100 mM HEPES, pH 7.5, 1.6 M ammonium sulfate, 7-9% (v/v) glycerol Synechocystis sp.

Protein Variants

EC Number Protein Variants Comment Organism
3.1.3.11 D198H more than 10fold reduction in kcat value, reduction in IC50 value of AMP Synechocystis sp.
3.1.3.11 D200A simultaneous decrease in substrate affinity and kcat value Synechocystis sp.
3.1.3.11 D97A almost complete loss of activity Synechocystis sp.
3.1.3.11 E100A almost complete loss of activity Synechocystis sp.
3.1.3.11 E225A almost complete loss of activity Synechocystis sp.
3.1.3.11 E57A almost complete loss of activity Synechocystis sp.
3.1.3.11 F309A greatly reduced activity Synechocystis sp.
3.1.3.11 H215A greatly reduced activity Synechocystis sp.
3.1.3.11 K134A greatly reduced activity, reduction in IC50 value of AMP Synechocystis sp.
3.1.3.11 K29A 2fold increase in activity Synechocystis sp.
3.1.3.11 N213A greatly reduced activity Synechocystis sp.
3.1.3.11 R164A greatly reduced activity Synechocystis sp.
3.1.3.11 R176A simultaneous decrease in substrate affinity and kcat value Synechocystis sp.
3.1.3.11 R178G greatly reduced activity Synechocystis sp.
3.1.3.11 R307A greatly reduced activity Synechocystis sp.
3.1.3.11 R314A greatly reduced activity Synechocystis sp.
3.1.3.11 T102A more than 10fold reduction in kcat value, reduction in IC50 value of AMP Synechocystis sp.
3.1.3.11 Y131A greatly reduced activity Synechocystis sp.
3.1.3.37 D198A inactive Synechocystis sp.
3.1.3.37 D200A inactive Synechocystis sp.
3.1.3.37 D33A inactive Synechocystis sp.
3.1.3.37 D97A inactive Synechocystis sp.
3.1.3.37 E100A inactive Synechocystis sp.
3.1.3.37 E225A inactive Synechocystis sp.
3.1.3.37 E57A inactive Synechocystis sp.
3.1.3.37 K134A inactive Synechocystis sp.
3.1.3.37 K29A the mutant shows increased activity compared to the wild type enzyme Synechocystis sp.
3.1.3.37 R176A inactive Synechocystis sp.
3.1.3.37 R178G the mutant shows very low activity compared to the wild type enzyme Synechocystis sp.
3.1.3.37 T102A inactive Synechocystis sp.
3.1.3.37 Y131A inactive Synechocystis sp.

Inhibitors

EC Number Inhibitors Comment Organism Structure
3.1.3.11 AMP
-
 Synechocystis sp.

KM Value [mM]

EC Number KM Value [mM] KM Value Maximum [mM] Substrate Comment Organism Structure
3.1.3.11 0.08
-
 D-fructose 1,6-bisphosphate mutant N213A, Hill coefficient 1.8, pH 8.0, 28°C Synechocystis sp.
3.1.3.11 0.08
-
 D-fructose 1,6-bisphosphate mutant R314A, Hill coefficient 1.8, pH 8.0, 28°C Synechocystis sp.
3.1.3.11 0.08
-
 D-fructose 1,6-bisphosphate wild-type, Hill coefficient 1.9, pH 8.0, 28°C Synechocystis sp.
3.1.3.11 0.1
-
 D-fructose 1,6-bisphosphate mutant H215A, Hill coefficient 1.4, pH 8.0, 28°C Synechocystis sp.
3.1.3.11 0.1
-
 D-fructose 1,6-bisphosphate mutant T102A, Hill coefficient 2.5, pH 8.0, 28°C Synechocystis sp.
3.1.3.11 0.11
-
 D-fructose 1,6-bisphosphate mutant R164A, Hill coefficient 2.1, pH 8.0, 28°C Synechocystis sp.
3.1.3.11 0.15
-
 D-fructose 1,6-bisphosphate mutant K29A, Hill coefficient 2.0, pH 8.0, 28°C Synechocystis sp.
3.1.3.11 0.17
-
 D-fructose 1,6-bisphosphate mutant D198H, Hill coefficient 1.9, pH 8.0, 28°C Synechocystis sp.
3.1.3.11 0.21
-
 D-fructose 1,6-bisphosphate mutant F309A, Hill coefficient 1.8, pH 8.0, 28°C Synechocystis sp.
3.1.3.11 0.24
-
 D-fructose 1,6-bisphosphate mutant R307A, Hill coefficient 1.6, pH 8.0, 28°C Synechocystis sp.
3.1.3.11 0.26
-
 D-fructose 1,6-bisphosphate mutant Y131A, Hill coefficient 1.6, pH 8.0, 28°C Synechocystis sp.
3.1.3.11 0.5
-
 D-fructose 1,6-bisphosphate mutant K134A, Hill coefficient 2.2, pH 8.0, 28°C Synechocystis sp.
3.1.3.11 0.61
-
 D-fructose 1,6-bisphosphate mutant R178A, Hill coefficient 1.1, pH 8.0, 28°C Synechocystis sp.
3.1.3.11 1.32
-
 D-fructose 1,6-bisphosphate mutant D200A, Hill coefficient 1.4, pH 8.0, 28°C Synechocystis sp.
3.1.3.11 3.2
-
 D-fructose 1,6-bisphosphate mutant R176A, Hill coefficient 1.2, pH 8.0, 28°C Synechocystis sp.
3.1.3.37 0.24
-
 sedoheptulose 1,7-bisphosphate in 50 mM Tris/HCl, pH 8.0, 15 mM MgCl2, 10 mM dithiothreitol, at 289°C Synechocystis sp.

Natural Substrates/ Products (Substrates)

EC Number Natural Substrates Organism Comment (Nat. Sub.) Natural Products Comment (Nat. Pro.) Rev. Reac.
3.1.3.37 sedoheptulose 1,7-bisphosphate + H2O Synechocystis sp.
-
 sedoheptulose 7-phosphate + phosphate
-
 ?

Organism

EC Number Organism UniProt Comment Textmining
3.1.3.11 Synechocystis sp. P73922
-
-
3.1.3.37 Synechocystis sp. P73922
-
-

Purification (Commentary)

EC Number Purification (Comment) Organism
3.1.3.37 Ni2+-NTA resin column chromatography Synechocystis sp.

Substrates and Products (Substrate)

EC Number Substrates Comment Substrates Organism Products Comment (Products) Rev. Reac.
3.1.3.11 D-fructose 1,6-bisphosphate + H2O
-
 Synechocystis sp. D-fructose 6-phosphate + phosphate
-
 ?
3.1.3.37 sedoheptulose 1,7-bisphosphate + H2O
-
 Synechocystis sp. sedoheptulose 7-phosphate + phosphate
-
 ?

Synonyms

EC Number Synonyms Comment Organism
3.1.3.11 slr2094
-
 Synechocystis sp.
3.1.3.37 FBP/SBPase
-
 Synechocystis sp.
3.1.3.37 fructose-1,6/sedoheptulose-1,7-bisphosphatase
-
 Synechocystis sp.

Turnover Number [1/s]

EC Number Turnover Number Minimum [1/s] Turnover Number Maximum [1/s] Substrate Comment Organism Structure
3.1.3.11 0.63
-
 D-fructose 1,6-bisphosphate mutant D200A, Hill coefficient 1.4, pH 8.0, 28°C Synechocystis sp.
3.1.3.11 0.7
-
 D-fructose 1,6-bisphosphate mutant R176A, Hill coefficient 1.2, pH 8.0, 28°C Synechocystis sp.
3.1.3.11 0.7
-
 D-fructose 1,6-bisphosphate mutant T102A, Hill coefficient 2.5, pH 8.0, 28°C Synechocystis sp.
3.1.3.11 0.84
-
 D-fructose 1,6-bisphosphate mutant D198H, Hill coefficient 1.9, pH 8.0, 28°C Synechocystis sp.
3.1.3.11 1.2
-
 D-fructose 1,6-bisphosphate mutant H215A, Hill coefficient 1.4, pH 8.0, 28°C Synechocystis sp.
3.1.3.11 1.2
-
 D-fructose 1,6-bisphosphate mutant Y131A, Hill coefficient 1.6, pH 8.0, 28°C Synechocystis sp.
3.1.3.11 1.8
-
 D-fructose 1,6-bisphosphate mutant K134A, Hill coefficient 2.2, pH 8.0, 28°C Synechocystis sp.
3.1.3.11 2.3
-
 D-fructose 1,6-bisphosphate mutant R164A, Hill coefficient 2.1, pH 8.0, 28°C Synechocystis sp.
3.1.3.11 2.6
-
 D-fructose 1,6-bisphosphate mutant R314A, Hill coefficient 1.8, pH 8.0, 28°C Synechocystis sp.
3.1.3.11 5.2
-
 D-fructose 1,6-bisphosphate mutant F309A, Hill coefficient 1.8, pH 8.0, 28°C Synechocystis sp.
3.1.3.11 5.2
-
 D-fructose 1,6-bisphosphate mutant N213A, Hill coefficient 1.8, pH 8.0, 28°C Synechocystis sp.
3.1.3.11 5.2
-
 D-fructose 1,6-bisphosphate mutant R307A, Hill coefficient 1.6, pH 8.0, 28°C Synechocystis sp.
3.1.3.11 7.1
-
 D-fructose 1,6-bisphosphate mutant R178A, Hill coefficient 1.1, pH 8.0, 28°C Synechocystis sp.
3.1.3.11 10.5
-
 D-fructose 1,6-bisphosphate wild-type, Hill coefficient 1.9, pH 8.0, 28°C Synechocystis sp.
3.1.3.11 21.3
-
 D-fructose 1,6-bisphosphate mutant K29A, Hill coefficient 2.0, pH 8.0, 28°C Synechocystis sp.
3.1.3.37 4.2
-
 sedoheptulose 1,7-bisphosphate in 50 mM Tris/HCl, pH 8.0, 15 mM MgCl2, 10 mM dithiothreitol, at 289°C Synechocystis sp.

IC50 Value

EC Number IC50 Value IC50 Value Maximum Comment Organism Inhibitor Structure
3.1.3.11 0.015
-
 mutant Y131A, pH 8.0, 28°C Synechocystis sp. AMP
3.1.3.11 0.0214
-
 mutant R176A, pH 8.0, 28°C Synechocystis sp. AMP
3.1.3.11 0.027
-
 mutant R178A, pH 8.0, 28°C Synechocystis sp. AMP
3.1.3.11 0.028
-
 mutant K29A, pH 8.0, 28°C Synechocystis sp. AMP
3.1.3.11 0.034
-
 wild-type, pH 8.0, 28°C Synechocystis sp. AMP
3.1.3.11 0.21
-
 mutant N213A, pH 8.0, 28°C Synechocystis sp. AMP
3.1.3.11 0.225
-
 mutant D200A, pH 8.0, 28°C Synechocystis sp. AMP
3.1.3.11 1.23
-
 mutant D198H, pH 8.0, 28°C Synechocystis sp. AMP
3.1.3.11 1.241
-
 mutant T102A, pH 8.0, 28°C Synechocystis sp. AMP
3.1.3.11 2.099
-
 mutant K134A, pH 8.0, 28°C Synechocystis sp. AMP

kcat/KM [mM/s]

EC Number kcat/KM Value [1/mMs-1] kcat/KM Value Maximum [1/mMs-1] Substrate Comment Organism Structure
3.1.3.11 0.21
-
 D-fructose 1,6-bisphosphate mutant R176A, pH 8.0, 28°C Synechocystis sp.
3.1.3.11 0.48
-
 D-fructose 1,6-bisphosphate mutant D200A, pH 8.0, 28°C Synechocystis sp.
3.1.3.11 3.6
-
 D-fructose 1,6-bisphosphate mutant K134A, pH 8.0, 28°C Synechocystis sp.
3.1.3.11 4.7
-
 D-fructose 1,6-bisphosphate mutant Y131A, pH 8.0, 28°C Synechocystis sp.
3.1.3.11 4.8
-
 D-fructose 1,6-bisphosphate mutant D198H, pH 8.0, 28°C Synechocystis sp.
3.1.3.11 6.8
-
 D-fructose 1,6-bisphosphate mutant T102A, pH 8.0, 28°C Synechocystis sp.
3.1.3.11 12
-
 D-fructose 1,6-bisphosphate mutant H215A, pH 8.0, 28°C Synechocystis sp.
3.1.3.11 12
-
 D-fructose 1,6-bisphosphate mutant R178A, pH 8.0, 28°C Synechocystis sp.
3.1.3.11 22
-
 D-fructose 1,6-bisphosphate mutant R164A, pH 8.0, 28°C Synechocystis sp.
3.1.3.11 22
-
 D-fructose 1,6-bisphosphate mutant R307A, pH 8.0, 28°C Synechocystis sp.
3.1.3.11 25
-
 D-fructose 1,6-bisphosphate mutant F309A, pH 8.0, 28°C Synechocystis sp.
3.1.3.11 32
-
 D-fructose 1,6-bisphosphate mutant R314A, pH 8.0, 28°C Synechocystis sp.
3.1.3.11 61
-
 D-fructose 1,6-bisphosphate mutant N213A, pH 8.0, 28°C Synechocystis sp.
3.1.3.11 130
-
 D-fructose 1,6-bisphosphate wild-type, pH 8.0, 28°C Synechocystis sp.
3.1.3.11 140
-
 D-fructose 1,6-bisphosphate mutant K29A, pH 8.0, 28°C Synechocystis sp.
3.1.3.37 17.5
-
 sedoheptulose 1,7-bisphosphate in 50 mM Tris/HCl, pH 8.0, 15 mM MgCl2, 10 mM dithiothreitol, at 289°C Synechocystis sp.